Improved traceability of Shiga-toxin-producing <Emphasis Type="Italic">Escherichia coli</Emphasis> using CRISPRs for detection and typing

Among strains of Shiga-toxin-producing Escherichia coli (STEC), seven serogroups (O26, O45, O103, O111, O121, O145, and O157) are frequently associated with severe clinical illness in humans. The development of methods for their reliable detection from complex samples such as food has been challenging thus far, and is currently based on the PCR detection of the major virulence genes stx1, stx2, and eae, and O-serogroup-specific genes. However, this approach lacks resolution. Moreover, new STEC serotypes are continuously emerging worldwide. For example, in May 2011, strains belonging to the hitherto rarely detected STEC serotype O104:H4 were identified as causative agents of one of the world’s largest outbreak of disease with a high incidence of hemorrhagic colitis and hemolytic uremic syndrome in the infected patients. Discriminant typing of pathogens is crucial for epidemiological surveillance and investigations of outbreaks, and especially for tracking and tracing in case of accidental and deliberate contamination of food and water samples. Clustered regularly interspaced short palindromic repeats (CRISPRs) are composed of short, highly conserved DNA repeats separated by unique sequences of similar length. This distinctive sequence signature of CRISPRs can be used for strain typing in several bacterial species including STEC. This review discusses how CRISPRs have recently been used for STEC identification and typing.     

Bench-scale composting of source-separated human faeces for sanitation

In urine-diverting toilets, urine and faeces are collected separately so that nutrient content can be recycled unmixed. Faeces should be sanitized before use in agriculture fields due to the presence of possible enteric pathogens. Composting of human faeces with food waste was evaluated as a possible method for this treatment. Temperatures were monitored in three 78-L wooden compost reactors fed with faeces-to-food waste substrates (F:FW) in wet weight ratios of 1:0, 3:1 and 1:1, which were observed for approximately 20 days. To achieve temperatures higher than 15 degrees C above ambient, insulation was required for the reactors. Use of 25-mm thick styrofoam insulation around the entire exterior of the compost reactors and turning of the compost twice a week resulted in sanitizing temperatures (>or=50 degrees C) to be maintained for 8 days in the F:FW=1:1 compost and for 4 days in the F:FW=3:1 compost. In these composts, a reduction of >3 log(10) for E. coli and >4 log(10) for Enterococcus spp. was achieved. The F:FW=1:0 compost, which did not maintain >or=50 degrees C for a sufficiently long period, was not sanitized, as the counts of E. coli and Enterococcus spp. increased between days 11 and 15. This research provides useful information on the design and operation of family-size compost units for the treatment of source-separated faeces and starchy food residues, most likely available amongst the less affluent rural/urban society in Uganda.     

Water quality assessment of Almatti Reservoir of Bijapur (Karnataka State, India) with special reference to zooplankton

The present investigation deals with the limnobiotic status of Almatti reservoir from February, 2003 to January, 2005. The study revealed that, the distribution and population density of zooplankton species depend upon the physico-chemical factors of the environment. Statistical analysis showed that there exists a significant relation between the biological and non-biological factors. The benthic fauna constituting the food of fish can be utilized for extensive culture operation so that the nutrients in the reservoir are not only properly cycled but also serve as a check on further eutrophication.     

Nanotechnology, risk and the environment: a review

Nanotechnologies are already interacting with the environment. Scientists and engineers are manipulating matter at the nanoscale, and these nanoscale processes and products are being used by industry in commercially available products. These products are either applied directly to the environment or end up in the environment through indirect pathways. This review examines the state of current environmental risk assessment of nanotechnologies. Nanotechnology is described generally, then both the possible benefits of nanotechnology and the risks are reviewed in a traditional way. Subsequently, a philosophical criticism of the traditional way of looking at risks is offered.     

Effects of environmental methylmercury on the health of wild birds, mammals, and fish

Wild piscivorous fish, mammals, and birds may be at risk for elevated dietary methylmercury intake and toxicity. In controlled feeding studies, the consumption of diets that contained Hg (as methylmercury) at environmentally realistic concentrations resulted in a range of toxic effects in fish, birds, and mammals, including behavioral, neurochemical, hormonal, and reproductive changes. Limited field-based studies, especially with certain wild piscivorous bird species, e.g., the common loon, corroborated laboratory-based results, demonstrating significant relations between methylmercury exposure and various indicators of methylmercury toxicity, including reproductive impairment. Potential population effects in fish and wildlife resulting from dietary methylmercury exposure are expected to vary as a function of species life history, as well as regional differences in fish-Hg concentrations, which, in turn, are influenced by differences in Hg deposition and environmental methylation rates. However, population modeling suggests that reductions in Hg emissions could have substantial benefits for some common loon populations that are currently experiencing elevated methylmercury exposure. Predicted benefits would be mediated primarily through improved hatching success and development of hatchlings to maturity as Hg concentrations in prey fish decline. Other piscivorous species may also benefit from decreased Hg exposure but have not been as extensively studied as the common loon.     

The Bias of Complementarity Hotspots toward Marginal Populations

Abstract: It has been suggested that using complementarity to identify networks of important areas for conserving biodiversity may preferentially select areas within the margins of species ranges. We tested this idea by examining the location of complementarity hotspots in relation to two measures of range core-periphery. The first measures patterns of aggregation among records within each species' range to identify areas within the core (i.e., areas with aggregated distributions) and periphery (i.e., areas with scattered distributions) of the range. The second measures spatial turnover among species to identify areas with a high density of range edges. For three selected groups of terrestrial vertebrates in Europe—mammals, birds, and herptiles—areas chosen based on complementarity were located within the margins of species' ranges more often than expected by chance. This pattern was consistent for the two measures of core-periphery we used. The bias of complementarity hotspots toward marginal populations is especially important for species with restricted range sizes. If extinctions are determined mainly by demographic factors, then selecting areas at the peripheries of species' ranges might be a poor option. But if extinctions are determined mainly by extrinsic factors, then peripheral populations might be important to ensure the long-term persistence of species.     

Molecular response of the sponge Suberites domuncula to bacterial infection

The aim of this study was the documentation of the molecular immune response of Suberites domuncula upon bacterial infection. Additionally, the bacteria that are naturally present in the sponge after prolonged aquarium maintenance were characterized. After 6 months of maintenance of S. domuncula in seawater aquaria, only one bacterial 16S rDNA sequence could be recovered, which belongs to the genus Pseudomonas. Concomitantly, morphologically uniform bacteria were found encapsulated in bacteriocytes. These findings indicate that certain bacteria, possibly of the genus Pseudomonas, are able to persist for long periods in host bacteriocytes. Subsequent to performing a previously established infection assay with S. domuncula, a potentially pathogenic Vibrio sp. was isolated from the tissues. Furthermore, the host tissue disintegrated and asexual propagation bodies (gemmules) were formed. In order to gain insights into the molecular events occurring after bacterial infection, the stress-response kinases, p38 protein kinase and JNK protein kinase, were analyzed. It is demonstrated that these two kinases are activated (phosphorylated) upon incubation of the tissue with the bacterial endotoxin lipopolysaccharide (LPS). Moreover, LPS strongly inhibits protein synthesis. It is concluded that there are many functionally different interactions between S. domuncula and bacteria and that the animal possesses mechanisms to differentiate between bacteria and to respond accordingly.     

Genetic, morphological, and chemical divergence in the sponge genus Latrunculia (Porifera: Demospongiae) from New Zealand

We undertook a comprehensive study of Latrunculia in New Zealand to determine the relationship between taxonomic, environmental, and chemical variation within the genus. Sponges were collected from five locations around New Zealand: Three Kings Islands, Tutukaka, Wellington, Kaikoura, and Doubtful Sound. Allozyme electrophoresis at nine polymorphic loci indicated that sponges from each geographic location were genetically distinct, and that they displayed genetic differences of the magnitude usually associated with reproductively isolated species (Nei's D between locations =0.375-2.476). Additionally, the comparisons revealed that the green and brown colour morphs of Latrunculia that are sympatric at Three Kings Islands and Kaikoura are distinct from each other, and that there are two genetic groups within the green sponges in Doubtful Sound. On the basis of genetic data we conclude that there are at least eight species of Latrunculia in New Zealand waters, not one to four as had been previously thought. Morphological comparisons of the eight genetic species based on skeletal characters (i.e. skeletal organisation of the choanosome, spicule composition, size, and geometry) indicated that the eight Latrunculia species fell into only two morphological groups that could be easily diagnosed on the basis of discorhabd type. Within these two primary morphological groups, skeletal characteristics among the eight species largely overlap and are not diagnostic. These findings emphasise the limitations of traditional taxonomic methods based solely on skeletal characters for distinguishing species of Latrunculia. However, multivariate analysis (MANOVA and CDA) based on six measured skeletal variables did reveal significant morphological variation among the species (Pillai's Trace=3.28, F=6.90, P<0.0001), supporting the division of the genus into eight species. Comparisons of chemical extracts from Latrunculia also showed that the amounts of five different bioactive compounds (discorhabdins A, B, C, D, and J) varied predictably among the eight species. This finding suggests that discorhabdin variation within Latrunculia, previously thought to be associated with intra-specific environmental variability, is more likely to reflect differences among species rather than phenotypic plasticity. Our results also highlight the importance of thorough taxonomic studies associated with marine natural products research to understand fully the variation in bioactive properties among individuals. The potential processes underlying the unusually high speciation rates in New Zealand Latrunculia that are indicated in our study are discussed.